Guanine quadruplex structures (GQSs) play important roles in the regulation of gene expression and cellular processes. Recent studies provide strong evidence for the formation and function of DNA and RNA GQSs in human cells. However, whether GQSs form and are functional in plants remains essentially unexplored. On the basis of circular dichroism (CD)-detected titration, UV-detected melting, in-line probing (ILP) and reporter gene assay studies, we report the first example of a plant RNA GQS that inhibits translation. This GQS is located within the 5′-UTR of the ATAXIA TELANGIECTASIA-MUTATED AND RAD3-RELATED (ATR) mRNA of Arabidopsis thaliana (mouse-ear cress). We show that this GQS is highly stable and is thermodynamically favoured over a competing hairpin structure in the 5′-UTR at physiological K+ and Mg2+ concentrations. Results from ILP reveal the secondary structure of the RNA and support formation of the GQS in vitro in the context of the complete 5′-UTR. Transient reporter gene assays performed in living plants reveal that the GQS inhibits translation but not transcription, implicating this GQS as a translational repressor in vivo. Our results provide the first complete demonstration of the formation and function of a regulatory RNA GQS in plants and open new avenues to explore potential functional roles of GQS in the plant kingdom.
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Research Article|
March 20 2015
A stable RNA G-quadruplex within the 5′-UTR of Arabidopsis thaliana ATR mRNA inhibits translation
Chun Kit Kwok;
*Department of Chemistry, Pennsylvania State University, University Park, PA 16802, U.S.A.
†Center for RNA Molecular Biology, Pennsylvania State University, University Park, PA 16802, U.S.A.
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Yiliang Ding;
*Department of Chemistry, Pennsylvania State University, University Park, PA 16802, U.S.A.
†Center for RNA Molecular Biology, Pennsylvania State University, University Park, PA 16802, U.S.A.
‡Department of Biology, Pennsylvania State University, University Park, PA 16802, U.S.A.
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Saima Shahid;
Saima Shahid
†Center for RNA Molecular Biology, Pennsylvania State University, University Park, PA 16802, U.S.A.
‡Department of Biology, Pennsylvania State University, University Park, PA 16802, U.S.A.
§Plant Biology Graduate Program, Pennsylvania State University, University Park, PA 16802, U.S.A.
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Sarah M. Assmann;
Sarah M. Assmann
4
†Center for RNA Molecular Biology, Pennsylvania State University, University Park, PA 16802, U.S.A.
‡Department of Biology, Pennsylvania State University, University Park, PA 16802, U.S.A.
§Plant Biology Graduate Program, Pennsylvania State University, University Park, PA 16802, U.S.A.
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Philip C. Bevilacqua
Philip C. Bevilacqua
4
*Department of Chemistry, Pennsylvania State University, University Park, PA 16802, U.S.A.
†Center for RNA Molecular Biology, Pennsylvania State University, University Park, PA 16802, U.S.A.
§Plant Biology Graduate Program, Pennsylvania State University, University Park, PA 16802, U.S.A.
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Publisher: Portland Press Ltd
Received:
August 14 2014
Revision Received:
December 22 2014
Accepted:
January 26 2015
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2015 Biochemical Society
2015
Biochem J (2015) 467 (1): 91–102.
Article history
Received:
August 14 2014
Revision Received:
December 22 2014
Accepted:
January 26 2015
Citation
Chun Kit Kwok, Yiliang Ding, Saima Shahid, Sarah M. Assmann, Philip C. Bevilacqua; A stable RNA G-quadruplex within the 5′-UTR of Arabidopsis thaliana ATR mRNA inhibits translation. Biochem J 1 April 2015; 467 (1): 91–102. doi: https://doi.org/10.1042/BJ20141063
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